类风湿关节炎关节液中抗环瓜氨酸肽抗体的测定及临床意义

目的 检测类风湿关节炎(RA)患者关节液中抗环瓜氨酸肽(CCP)抗体,了解抗CCP抗体在RA关节液中的阳性率及对RA的临床意义.方法 应用酶联免疫吸附测定法检测43例RA患者关节液抗CCP抗体和对照组21例患者的关节液抗CCP抗体,并测定RA患者的一些临床及实验室指标.结果 43例RA患者中,关节液抗CCP抗体阳性的32例,阴性的11例.对照组2l例患者,其中4例UA关节液抗CCP抗体阳性,其余均呈阴性.RA组关节液抗CCP抗体的阳性率高于对照组,74.4%VS 19.0%(χ2=17.578,P<0.01).RA关节液抗CCP抗体阳性组与阴性组相比较,在年龄,病程和晨僵方面差异均无统计学意义,而关节液抗CCP抗体阳性组血沉(ESR)、C反应蛋白(CRP)和DAS 28评分均比阴性组高,ESR(80.43±20.18)mm/1 h VS(41.22±9.56)mm/1 h、CRP(39.88±10.30)mg/L VS(19.29±2.97)mg/L、DAS 28(5.70±0.51)分vs(4.90±0.70)分(均P<0.05),抗CCP抗体与ESR和CRP之间呈正相关(r=0.250,P<0.05,r=0.450,P<0.01).结论 RA关节液抗CCP抗体对RA的早期诊断及对疾病活动的判断有重要意义.     

类风湿关节炎患者滑膜组织中瓜氨酸化蛋白质及肽酰基精氨酸脱亚氨酶4的表达研究

目的 研究瓜氨酸化蛋白质(citrullinated protein,Cit P)及肽酰基精氨酸脱亚氨酶4(peptidylarginine deiminase,PAD4)在类风湿关节炎(RA)患者关节滑膜组织(synovial tissue,ST)中的表达。方法 无菌条件下获取手术切除的RA和骨性关节炎(osteoarthritis,OA)患者的关节ST,用免疫组织化学法检测ST中Cit P、Ig G及补体C3成分;western blot法检测RA和OA患者ST中PAD4蛋白的表达情况。分析RA患者ST中PAD4蛋白质表达与血清抗环瓜氨酸肽(CCP)抗体、抗链球菌溶血素O(ASO)、类风湿因子(RF)、C反应蛋白(CRP)和疾病活动性评分系统(DAS28)等临床指标的相关性。结果 RA和OA患者ST中均有Ig G和C3表达,但Cit P仅在RA患者ST中表达;RA患者ST中PAD4蛋白的表达量明显高于OA患者(P<0.05),且其表达量与CRP存在负相关关系(P<0.05),但未发现与其他临床指标存在相关性。结论 RA患者ST中PAD4、Cit P蛋白表达水平较高,PAD4在RA的致病过程中可能发挥一定的作用。     

类风湿性关节炎滑膜组织PADI4的表达

目的 研究类风湿关节炎(RA)、骨关节炎(OA)和强直性脊柱炎(AS)滑膜液中肽酰基精氧酸脱亚胺酶4(PADI4)的表达.方法 收集RA(n=73)、OA(n=96)和AS(n=32)滑膜液标本及RA(n=6)、OA(n=6)和AS(n=32)关节滑膜标本,应用酶联免疫吸附法(ELISA)和蛋白印迹分析的方法检测滑膜液中PADI4蛋白表达,应用实时定量PCR比较RA、OA和AS患者滑膜中PADI4的mRNA的表达,并研究抗环瓜氨酸肽(CCP)抗体和类风湿因子(RF)、C反应蛋白(CRP)与PADI4的相关性.结果 RA患者滑膜液中PADI4、抗CCP抗体、RF含量较高,OA、AS患者滑膜液中PADI4、抗CCP抗体、RF含量无明显改变,RA患者与OA、AS患者比较差异有统计学意义(P<0.05).结论 以上结果证实RA患者滑膜中PADI4表达较高,PADI4在RA发病过程中发挥作用,PADI4的表达水平可能与一些临床表现有关.     

Dynamic changes in cytokine levels in serum and synovial fluid following filtration leukocytapheresis therapy in patients with rheumatoid arthritis

We attempted to determine whether various cytokine levels in the serum and synovial fluid (SF) of rheumatoid arthritis (RA) patients are influenced by the performance of filtration leukocytapheresis (LCP). The filtration LCP procedure that used a Cellsorba® column (LCP group: n=22; responder subgroup: n=17, non‐responder subgroup: n=5) or sham apheresis (control group; n=7) was repeated three times at 1‐week intervals. Serum (LCP group, n=22; control group, n=7) and SF (LCP group, n=6; control group, n=3) samples were collected before and after LCP. Levels of tumor necrosis factor α (TNFα), interleukins (IL‐1β, IL–2, IL‐6, IL‐8, IL‐10, and IL‐15), granulocyte–macrophage colony–stimulating factor (GM‐CSF), monocyte chemoattractant protein–1 (MCP‐1), RANTES were measured by an enzyme‐linked immunosorbent assay. Serum TNFα, IL‐15, and RANTES were significantly reduced only in the LCP group. Serum IL‐10 significantly increased only in the LCP group. In the LCP subgroup, serum IL‐15, GM‐CSF, and RANTES levels were reduced significantly, while serum IL‐10 levels increased significantly only in the responder group after treatment. Serum TNFα levels were reduced significantly in both subgroups. Changes in serum IL‐10 correlated positively with the improvement of patient's assessment of pain and global severity, and physician's assessment of global severity. These results indicate that the removal of leukocytes from the peripheral blood of RA patients provokes dynamic changes in some cytokine levels in the serum and/or synovial fluid. These changes may explain some of the mechanisms by which the articular symptoms are improved by filtration LCP. J. Clin. Apheresis. 16:74–81, 2001. © 2001 Wiley‐Liss, Inc.     

关节液各检验指标与关节炎类型的相关性

目的找到关节液中各检验指标的改变与不同病因之间的相关性。方法将关节炎病例按常见类型分为6组,即骨性关节炎、反应性关节炎、痛风性关节炎、类风湿关节炎、创伤性关节炎、其他类型关节炎。每组随机选择37例患者,抽取关节液,并检测关节液中白细胞(WBC)、总蛋白(TP)、清蛋白(ALB)、糖(GLU)、尿酸(UA)、抗链球菌溶血素O(ASO)、类风湿因子(RF)、C反应蛋白(CRP)、免疫球蛋白G(IgG)、免疫球蛋白A(IgA)、免疫球蛋白M(IgM)、补体C3、补体C4、白细胞介素(IL)-1β、IL-6、肿瘤坏死因子-α(TNF-α)的水平,将检测数据进行分组统计。结果除CRP、IgG、IgA外,其余各测定指标(WBC、TP、ALB、GLU、UA、ASO、RF、IgM、C3、C4、IL-1β、IL-6、TNF-α)均存在差异。结论关节液中诸多成分的水平与病因相关。     

沙利度胺对类风湿关节炎滑膜成纤维样细胞影响的体外研究

目的研究沙利度胺(Thalidomide)对类风湿关节炎(RA)滑膜成纤维样细胞(FLS)体外培养时的增生殖分化特性的影响。方法关节镜、关节活检针取RA患者滑膜组织,分离培养和鉴定FLS,MTT法和反转录-聚合酶链反应(RT-PCR)方法观察沙利度胺对FLS细胞存活分数(SF)和c-fos、COX-2mRNA表达。结果RA-FLS体外培养呈良性增生。沙利度胺对RA-FLS的SF值的抑制作用最强(P<0.05)。生理剂量的沙利度胺干预时的浓度与FLS的SF值呈负相关。RA-FLS的c-fos、环氧化酶2(COX-2)mRNA表达率高,加入沙利度胺共孵育3天后c-fos和COX-2mRNA的表达率均明显下降。结论培养和鉴定RA滑膜细胞,证实体外培养传代的RA-FLS为非恶性无限制增生。实验剂量下沙利度胺通过抑制c-fos、COX-2的表达、降低FLS增殖能力等不同途径,发挥对RA滑膜细胞的免疫调节作用。     

Elevated soluble CD8 in the synovial fluid from patients with rheumatoid arthritis

Suppressor/cytotoxic T cells express the surface marker CD8, which can be measured in a soluble form in culture supernatants of activated human lymphocytes. Using a sandwich immunoassay, we assessed the levels of soluble CD8 (sCD8) in serum from patients with rheumatoid arthritis (RA; n = 82), patients with degenerative joint disease (DJD; n = 40), and healthy controls. There were no differences in serum sCD8 levels among these groups. In contrast, the levels of soluble CD8 in the synovial fluid (SF) from patients with RA (n = 53) were significantly increased compared with the levels in 23 samples from patients with DJD (821 +/- 110 U/ml versus 213 +/- 13 U/ml, p less than 0.001). Synovial fluid sCD8 levels in the RA group were strikingly elevated, to a maximum value of 5,026 U/ml. In the majority of RA SF specimens (39 of 53), the values were significantly higher in the SF than the serum. Although the RA group had higher values of sCD8, such values were not significantly correlated with measured laboratory or clinical parameters. Current clinical and laboratory methods of evaluating patients may not be adequate in dealing with the complexity and heterogeneity of RA. Soluble CD8 values may be useful in further grouping patients with this disease.     

Detectable levels of pyridinoline are present in synovial fluid from various patients with knee effusion: preliminary results*

Abstract. There is a major interest for using biochemical markers of bone metabolism as a non-invasive tool for diagnostic purposes in the field of bone and joint diseases. Based upon the fact that the pyridinium crosslinks of collagen are markers of bone and cartilage degradation, this study was designed to assess the presence of pyridinoline in synovial fluid samples originating from various arthritic and non-arthritic knee joints. Using a sample pooling method, significant levels of pyridinoline could be measured in synovial fluid by high performance liquid chromatography. Pyridinoline levels ranged from 19.3 ± 5.8 pmol mL^-1 (mean ± SD) in osteoarthritic knee joints up to 32.4 ± 14.6 pmol mL^-1 in rheumatoid arthritis joints. Pyridinoline levels in synovial fluid were not significantly correlated to disease duration and synovial fluid cell count, but were correlated to erythrocyte sedimentation rate in osteoarthritic patients (r= 0.99, P= 0.002). This study demonstrates that synovial fluid originating from knee effusion contains significant levels of pyridinoline which can be quantified by high performance liquid chromatography and could, therefore, be a tool to investigate the metabolism of a single joint.     

Protein oxidation markers in the serum and synovial fluid of psoriatic arthritis patients

The role of oxidative stress has been studied in rheumatoid arthritis (RA) and other inflammatory joint diseases to some extent, but its importance in psoriatic arthritis (PsA) has rarely been investigated. The aim of this study was to analyze the levels of protein oxidation markers, sulfhydryl (SH) and carbonyl (CO) groups, in the synovial fluid (SF) and serum of PsA patients and compare them with the findings in RA and osteoarthritis (OA) patients. A total of 49 subjects with a knee-joint effusion including 16 PsA, 18 RA, and 15 OA patients were studied. In all patients, the levels of SH groups measured in the serum and SF inversely correlated with the number of white blood cells (WBC) (P<0.05) and the percentage of polymorphonuclear leukocytes (PMN) (P<0.01) in SF. Serum SH levels inversely correlated with serum erythrocyte sedimentation rate (ESR) (P<0.02) and C-reactive protein (CRP) (P<0.05) values. The SH levels in SF were significantly lower in patients affected by PsA and RA compared to OA cases (P<0.02). The serum SH levels in PsA were lower than OA (P<0.001) and higher than RA patients (P<0.05). The serum and synovial levels of CO groups in PsA, RA, and OA patients were similar. Our study provides novel evidence on the involvement of protein oxidation in PsA and confirms the important role of oxidative stress in the pathogenesis of RA. These data suggest that antioxidant agents can potentially be a useful addition to the conventional therapy in the management of these diseases.     

类风湿关节炎患者RANTES及MCP-1的测定及其意义

目的 研究类风湿关节炎（RA）患者血清、关节滑液中调节活化正常T细胞表达和分泌因子（RANTES）及单桂细胞趋化蛋白1（MCP-1）表达水平,探讨其在RA致病中的作用。方法 用酶联免疫吸附法（ELISA）测定27例类风温关节炎患者血清及病变膝关节滑液中RANTES、MCP-1水平,井与外伤截肢者对照。结果 RA患者血清及滑液中RANTES、MCP-1水平明显高于时照组,且RA滑液中RANTES水平高于血清。结论 以上研究结果提示RA病变关节组织是RANTES产生的主要部位。     

实时定量PCR检测类风湿关节炎滑液单个核细胞MIP-1α mRNA的表达

目的了解类风湿关节炎（RA）患者滑液单个核细胞MIP-1α mRNA表达情况。方法用实时定量PCR技术检测8例活动期RA患者滑液和血清单个核细胞（SFMC和PBMC）中巨嗜细胞炎症蛋白-1α（MIP-1α）mRNA的表达水平。同时检测血清CRP和滑液白细胞数。结果RA患者SFMC MIP-1α mRNA表达水平比自身PBMC表达水平明显增高（P〈0.01），并且与血清CRP和滑液白细胞数呈正相关（r分别为0．9028和0．9120，P〈0．01）。结论 RA认患者可能通过SFMC高表达MIP-1α趋化炎性细胞向滑膜迁移，加重局部炎症反应。     

Collagenase-like (CL) peptidase activity in synovial fluid from patients with rheumatoid arthritis

We found the presence of collagenase-like (CL) peptidase in synovial fluid by a highly sensitive fluorescence assay using (succinyl-Gly-Pro-Leu-Gly-Pro)-4-methylcoumaryl-7-amide (Suc-GPLGP-MCA) as a substrate. Suc-GPLGP-MCA is hydrolyzed at the Leu-Gly bond by CL-peptidase. The CL-peptidase activity in synovial fluid was significantly higher in patients with rheumatoid arthritis (RA) than in patients with osteoarthritis (OA) and in arthropathy-free controls. No significant difference in CL-peptidase activity in synovial fluid was found between patients with OA and arthropathy-free controls.     

关节液中抗环瓜氨酸肽抗体在类风湿关节炎诊断和评估中的意义

目的:分析关节液中抗环瓜氨酸肽(CCP)抗体对类风湿关节炎(RA)的诊断和评估意义。方法:用ELISA法检测37例RA患者血清和关节液中的抗CCP抗体和13例对照组关节炎患者关节液中的抗CCP抗体,并测定类风湿因子(RF),记录RA患者的一些临床参数。对检测结果及临床参数进行统计分析。结果:关节液中抗CCP抗体对RA诊断的敏感度为72.9%,特异度为92.3%。RA组和对照关节炎组患者的关节液中抗CCP抗体的差异有显著性。RA组血清与关节液中抗CCP抗体的浓度高度相关(P<0.001),且关节液中抗CCP抗体与关节液中RF浓度有较高相关性(P<0.01)。RA组关节液中抗CCP抗体与病程相关,病程越长,抗CCP抗体滴度越高;而与肿胀关节数、压痛关节数、疾病活动评分(DAS28)、血清RF、红细胞沉降率(ESR)、C反应蛋白(CRP)和Steinbrocker放射线分级不相关。结论:关节液中抗CCP抗体对RA的诊断有高度特异性和较好敏感性,对其他关节炎有显著性差异,独立于一些临床参数,同时又与血清中抗CCP抗体及关节液中的RF相关。     

Synovial biopsies in inflammatory arthritis: precision medicine in rheumatoid arthritis

ABSTRACT

Introduction: Synovial tissue (ST) is composed of a lining and sublining layer and is the target tissue involved in the inflammatory arthritides (IA), in which there is lining layer hyperplasia, inflammatory cell influx, macrophage recruitment and change in number and behavior of lining fibroblasts. Understanding synovial pathology has been critical in providing insights into pathogenetic mechanisms of disease and therapeutics. Pathobiological insights into ST have been underpinned by progress in molecular analytic methods; research in this area holds promise in individualizing treatment and optimizing response.

Areas covered: We explore ST in IA and cover in-depth the utility of synovial biopsy and ST heterogeneity. We review recent advances in ST research and discuss implications with regards to therapeutic response. Finally, we provide perspectives on the identification of new drug targets and new diagnostic and prognostic markers.

Expert opinion: ST holds the potential to individualize therapy by detecting biomarkers of diagnosis, therapeutic choice, and treatment modification in IA. Advances in molecular biology including high-throughput omics are likely to provide information that has hitherto remained unknown. ST analyzes pre- and post-treatment needs to be standard of care; only by routinely collecting and analyzing ST will we achieve the precision medicine outcomes described herein.     

Comparative study of serum and synovial fluid interleukin-11 levels in patients with various arthritides

Objective: To determine the levels of serum and synovial fluid (SF) interleukin (IL)-11 in patients with various arthritides and estimate the contribution of IL-11 to acute phase response (APR).

Design and methods: Serum and SF IL-11 were measured by ELISA in patients with rheumatoid arthritis (RA, n = 31), seronegative spondyloarthritis (SSA, n = 23), gout (GT, n = 14) and osteoarthritis (OA, n = 20) and were correlated with ESR and acute phase proteins as well as with cytokines IL-1, IL-1β, IL-6, and TNF.

Results: IL-11 was detected in both serum and SF in each group, with IL-11 being statistically higher in SF than serum in all groups, suggesting reduced catabolism or increased synthesis of IL-11 intra-articularly. Median SF IL-11 levels were higher in OA patients than in other groups and in the treated than in the untreated RA subgroup. Moreover, serum and SF IL-11 were correlated significantly with each other, and moderately with the other cytokines examined in RA, SSA, and GT, but not in OA patients, while a significant negative correlation was found with a few of the inflammatory markers examined in each group.

Conclusions: Our findings provide evidence of extensive intra-articular expression of IL-11 in arthritides, especially in OA and treated RA patients, suggesting a protective role for IL-11 in joints, probably through the induction of tissue inhibitor of metalloproteinases.     

Leukocytapheresis in rheumatoid arthritis

In this article, we discussed leukocytapheresis (LCAP) for rheumatoid arthritis (RA). Recently, a simple and practical on-line continuous LACP system has been developed. It is equipped with a direct hemoperfusion column (Cellsorba^®, Asahikasei Medical Co., Ltd.) packed with fine-diameter polyester fibers, which are commonly used to adsorb white blood cells to prevent a graft-versus-host reaction during blood transfusion. Clinical trials revealed that LCAP is a effective and safe therapy for patients with drug-resistant RA or RA complicated with vasculitis. Because the procedure is simple and requires no plasma substitutes and the volume needed for extracorporeal circulation is less than that for other plasmapheresis, LCAP might be accepted as an optional therapeutic modality for active RA that was refractory to conventional drug therapy including biological agents. The mechanism of the efficiency of LCAP on RA is unclear. LCAP may cause a reduction of activated T cells from affected joints, down-regulation of Pgp on helper T cells and restoration of Treg function, and that may modify the abnormal cytokine balance. These findings may explain some of the mechanisms by which the articular symptoms are improved by LCAP.     

高场MRI扫描在手部类风湿性关节炎中的应用效果及对疾病活动度的评估价值

目的探讨高场磁共振成像(MRI)扫描在手部类风湿性关节炎(RA)中的应用效果及对疾病活动度的评估价值。方法选取2016年1月至2019年12月收治的93例手部RA患者作为研究对象,根据是否处于活动期将其分为活动期组(48例)与非活动期组(45例)。两组患者均行高场MRI扫描,比较两组的MRI征象、滑膜强化率及滑膜厚度。根据RA疾病活动度(DAS28评分)将活动期组患者分为轻、中、重度组,比较不同疾病活动度患者的MRI征象、滑膜强化率及滑膜厚度。结果活动期组的滑膜增厚、骨髓水肿、骨侵蚀、滑膜炎、腱鞘炎检出率、滑膜强化率及滑膜厚度均高于非活动期组,差异具有统计学意义(P<0.05)。重度组的滑膜增厚、骨髓水肿、骨侵蚀、滑膜炎、腱鞘炎检出率均高于轻、中度组,但差异无统计学意义(P>0.05)。重度组的滑膜强化率、滑膜厚度均优于轻、中度组,且中度组优于轻度组,差异具有统计学意义(P<0.05)。结论高场MRI扫描可清晰显示手部RA活动期病变特征,并能评估疾病活动度,为临床诊断RA提供参考依据。     

抗环瓜氨酸肽抗体联合类风湿因子诊断类风湿关节炎的临床意义

目的探讨类风湿关节炎(RA)血清标志物抗环瓜氨酸肽(CCP)抗体的临床价值,并分析其与类风湿因子(RF)的关联性。方法采用化学发光法检测CCP抗体,使用免疫散射比浊法检测RF,并对结果进行统计学分析。结果 113例RA患者抗CCP抗体对RA诊断的敏感性73.5%,阳性83例;84例非RA患者抗CCP抗体特异性97.6%,阳性2例。82例抗CCP抗体与RF检测结果相同(72.6%)。两者无相关性(P0.05)。抗CCP抗体与RF联合检测的敏感性84.1%,特异性98.8%。结论抗CCP抗体与RF抗体联合检测可提高RA的特异性。     

苦参碱对人成纤维样滑膜细胞生长的影响

目的：探讨苦参碱对类风湿关节炎的潜在治疗作用。方法原代培养人关节成纤维样滑膜细胞,传至第3～5代时,以0．4、0．8、1．2、1．6、2．0mg／mL等不同浓度的苦参碱处理细胞,分别于作用后24、48h用倒置显微镜观察细胞形态。采用四甲基偶氮唑蓝（MTT ）法检测不同浓度苦参碱处理的细胞生长情况,计算抑制率。结果不同浓度的苦参碱均可抑制人成纤维样滑膜细胞生长,MTT 检测结果显示细胞抑制率为2．52％～25．74％,随苦参碱药物浓度增加和处理时间延长,抑制作用增强。结论苦参碱可抑制人成纤维样滑膜细胞生长,可作为治疗类风湿关节炎的潜在药物。